Phosphorylation of synthetic peptides by a CDPK and plant SNF1-related protein kinase. Influence of proline and basic amino acid residues at selected positions

Jian Zhong Huang, Steven C Huber

Research output: Contribution to journalArticle

Abstract

Spinach (Spinacia oleracea L.) leaf sucrose-phosphate synthase (SPS) can be inactivated by phosphorylation of Ser-158 by calmodulin-like domain protein kinases (CDPKs) or SNF1-related protein kinases (SnRK1) in vitro. While the phosphorylation site sequence is relatively conserved, most of the deduced sequences of SPS from dicot species surrounding the Ser-158 regulatory phosphorylation site contain a Pro residue at P-4 (where P is the phosphorylated Ser); spinach is the exception and contains an Arg at P-4. We show that a Pro at P-4 selectively inhibits phosphorylation of the peptide by a CDPK relative to a SnRK1. The presence of a Pro at P-4, by allowing a tight turn in the peptide substrate, may interfere with proper binding of residues at P-5 and beyond. Both kinases had greater activity with peptides having basic residues at P-6 and P+5 (in addition to the known requirement for an Arg at P-3/P-4), and when the residue at P-6 was a His, the pH optimum for phosphorylation of the peptide was acid shifted. The results are used to predict proteins that may be selectively phosphorylated by SnRK1s (as opposed to CDPKs), such as SPS in dicot species, or may be phosphorylated in a pH-dependent manner.

Original languageEnglish (US)
Pages (from-to)1079-1087
Number of pages9
JournalPlant and Cell Physiology
Volume42
Issue number10
DOIs
StatePublished - Jan 1 2001

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Basic Amino Acids
sucrose-phosphate synthase
synthetic peptides
calmodulin
Calmodulin
Proline
protein kinases
Protein Kinases
proline
phosphorylation
Phosphorylation
Spinacia oleracea
Peptides
amino acids
peptides
Magnoliopsida
spinach
phosphotransferases (kinases)
Phosphotransferases
SNF1-related protein kinases

Keywords

  • CDPKs
  • Phosphorylation motif
  • Protein kinase specificity
  • SNF1-related protein kinase
  • Spinacia oleracea

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

Cite this

Phosphorylation of synthetic peptides by a CDPK and plant SNF1-related protein kinase. Influence of proline and basic amino acid residues at selected positions. / Huang, Jian Zhong; Huber, Steven C.

In: Plant and Cell Physiology, Vol. 42, No. 10, 01.01.2001, p. 1079-1087.

Research output: Contribution to journalArticle

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